scholarly journals 277 NUCLEAR STAGE AND p34cdc2 EXPRESSION IN DIFFERENT SIZES OF PREPUBERTAL GOAT OOCYTES

2005 ◽  
Vol 17 (2) ◽  
pp. 288
Author(s):  
B. Anguita ◽  
A.R. Jimenez-Macedo ◽  
D. Izquierdo ◽  
M.T. Paramio
Keyword(s):  
Developmental Competence ◽  
Rna Expression ◽  
Rt Pcr ◽  
Globin Mrna ◽  
Exact Test ◽  
Morphological Criteria ◽  
Before And After ◽  
Nuclear Stage ◽  
Cytoplasmic Maturation ◽  
Collection Time

Developmental competence of prepubertal (1- to 2-month old) goat oocytes is compromised, probably because of an incomplete cytoplasmic maturation. Oocyte selection for IVM-IVF is based on morphological criteria. The main regulator of oocyte nuclear maturation is maturation promoting factor (MPF), and it could also be involved in cytoplasmic maturation. The present study tried to determined p34cdc2 (the catalytic subunit of MPF) expression in oocytes of different sizes before and after IVM. Prepubertal goat oocytes were matured in a conventional IVM medium (TCM199 with serum, hormones, and cysteamine) for 27 h. At collection time, a sample of oocytes was classified into 4 groups according to their diameter (<110 μm; 110–125 μm; 125–135 μm; and >135 μm), and nuclear stage was evaluated. After IVM, oocytes were classified by diameter (as described before) and stained to analyze nuclear stage. Before and after IVM, a sample of 10 oocytes of each diameter group was frozen at −80°C. These oocytes were used to perform the detection of p34cdc2-RNA by RT-PCR. Briefly, RNA of 10 oocytes was extracted with TriReagent (Sigma-Aldrich, Madrid, Spain), and used to perform RT using the ThermoScript kit (Invitrogen, Barcelona, Spain). The cDNA corresponding to two oocytes was amplified by PCR, and each amplification band was measured by densitometry using the Quantity One PC program. Rabbit globin mRNA was used as an extrinsic control of the whole process. Nuclear stage data were analyzed by Fisher's exact test, and RT-PCR data were analyzed by one-way ANOVA test. At collection time, a high percentage of prepubertal goat oocytes had resumed meiosis. After IVM, the percentage of MII-oocytes was higher in larger-sized oocytes. At collection time, significantly higher p34cdc2-RNA expression was found in 125–135 μm oocytes. After IVM, no differences were found among oocyte groups. During maturation, a decrease of p34cdc2-RNA was found in 125–135 μm oocytes. In contrast, an increase of p34cdc2-RNA was found in 110-125 μm oocytes. The nuclear stage in the smallest oocytes show their reduced ability to resume meiosis. IVM-oocytes of different diameters showed no difference in p34cdc2-RNA expression. Table 1. p34cdc2-RNA expression of prepubertal goat oocytes according to nuclear stage and oocyte sizes This study was supported by MCYT (Spain), with the grant number AGL2000-0353.

scholarly journals ζ-, ε-, and γ-Globin mRNA in Blood Samples and CD71+ Cell Fractions from Fetuses and from Pregnant and Nonpregnant Women, with Special Attention to Identification of Fetal Erythroblasts

2001 ◽  
Vol 47 (4) ◽  
pp. 645-653 ◽  
Author(s):  
Anne Mette Høgh ◽  
Thomas Vauvert F Hviid ◽  
Britta Christensen ◽  
Steen Sørensen ◽  
Rasmus D Larsen ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Rt Pcr ◽  
Globin Mrna ◽  
Isolation And Identification ◽  
Blood Samples ◽  
Exact Test ◽  
Fisher's Exact Test ◽  
Significant Difference ◽  
Fisher’S Exact Test ◽  
Cell Fractions

Abstract Background: Information about the appearance of γ-, ε-, and ζ-globin mRNAs in fetal erythroblasts during gestation and about the presence and amounts of these mRNAs in pregnant and nonpregnant women is important from the perspective of using these molecules as a marker of fetal erythroblasts. A specific marker is necessary for isolation and identification of fetal nucleated red blood cells from maternal blood samples for use in antenatal diagnosis of fetal genetic or chromosomal abnormalities. Methods: We used a very sensitive reverse transcription-PCR (RT-PCR) method, coamplification analysis of γ- and ε-globin cDNA, and quantitative analysis of γ-globin mRNA based on competitive RT-PCR to investigate these aspects. Results: All adult whole-blood samples were negative for ε- and ζ-globin mRNA. Analyses of CD71+ cell fractions showed that specimens from 19 of 20 nonpregnant and 10 of 14 pregnant women (at 9–13 weeks of gestation) were positive for γ-globin mRNA (Fisher’s exact test, P = 0.13), and those from 3 of 20 nonpregnant and 5 of 14 pregnant women were positive for ζ-globin mRNA (Fisher’s exact test, P = 0.23). No ε-globin mRNA was detected in CD71+ cell fractions from 1-mL blood samples from adults. CD71+ cell fractions from eight fetal blood samples (at 17–20 weeks of gestation) were positive for all three globin mRNAs. We found no statistically significant difference between the amounts of γ-globin mRNA in pregnant and nonpregnant women. Conclusions: This study indicates that ε-globin mRNA might function as a marker for fetal CD71+ cells early in pregnancy. Although γ-globin mRNA can be detected in CD71+ cell fractions from most adults, these transcripts also may be of use because of a marked difference between adult and fetal values.

320 MPF ACTIVITY AND DEVELOPMENTAL COMPETENCE IN DIFFERENT SIZES OF PREPUBERTAL GOAT OOCYTES

2006 ◽  
Vol 18 (2) ◽  
pp. 267 ◽  
Author(s):  
B. Anguita ◽  
A. R. Jimenez-Macedo ◽  
D. Izquierdo ◽  
M. T. Paramio
Keyword(s):  
Embryonic Development ◽  
Kinase Activity ◽  
Developmental Competence ◽  
Kinase Assay ◽  
Oocyte Diameter ◽  
Cdc2 Kinase ◽  
Cell Extracts ◽  
Morphological Criteria ◽  
Blastocyst Rate ◽  
Cytoplasmic Maturation

Developmental competence of prepubertal goat oocytes recovered from a slaughterhouse is low, probably due to an incomplete cytoplasmic maturation. Regulation of cytoplasmic maturation is still unknown, although maturation-promoting factor (MPF) is suggested to play an important role in this process. To better understand the role of MPF in cytoplasmic maturation, we have studied MPF kinase activity in oocytes with different developmental competence. Ovaries were obtained from a slaughterhouse, and oocytes were recovered by slicing and were selected according to morphological criteria. Some oocytes were denuded and classified in diameter groups (<110 μm, 110–125 μm, 125–135 μm, and >135 μm), placed in lysis buffer (50 mM Tris-HCl (pH 7.5), 0.5 M NaCl, 5 mM EDTA, 0.01% Brij35, 1 mM phenyl methyl sultonyl fluoride (PMSF), 0.05 mg/mL leupeptin, 50 mM 2-mercaptoethanol, 25 mM α-glycerophosphate, 1 mM Na-orthovanadate) and frozen in liquid N2. Cell extracts were stored at −80°C until use. The rest of oocytes were matured in vitro in medium TCM199 supplemented with hormones, 10% (DBS), and 400 μM cysteamine, for 27 h in 5% CO2 in air and 38.5°C. After IVM, a sample of oocytes were also denuded, classified by diameters, and frozen as described above. The rest of oocytes were used for IVF in mDM with spermatozoa capacitated with heparin and ionomicin. After 24 h, presumptive zygotes were cultured for 7 days in medium SOF in 5% CO2, 5% O2, and 90% N2 at 38.5°C. At 48 h post-insemination, we added 0.1 μL FBS per embryo. Embryonic development was evaluated with Hoechst staining after IVC. MPF kinase activity was detected using the MESACUP cdc2 kinase assay kit (MBL Woburn, MA, USA). Briefly, the oocyte extract corresponding to 10 oocytes was mixed with 10× reaction buffer (25 mM HEPES buffer (pH 7.5), 10 mM MgCl2) and 10% biotinylated MV peptide (SLYSSPGGAYC). We added 0.1 mM ATP to start the reaction. The mixture was incubated at 30°C for 30 min. The reaction was finished by adding 200 μL of PBS containing 50 mM EGTA. The phosphorylated MV peptide was detected by specific antibody using an ELISA procedure, and the OD was measured at 492 nm. Fisher's exact test was used to analyze IVC results, and ANOVA to analyze cdc2 kinase activity results. We considered differences statistically significant when P < 0.05. Results are shown in Table 1. We observed that embryonic cleavage and blastocyst rate increased with oocyte diameter. The MPF activity detected was higher in the largest oocytes after IVM. As a consequence, we could establish that oocytes with a higher MPF activity are more capable of maintaining embryonic development until the blastocyst stage, which may indicate the important role that MPF plays in cytoplasmic maturation. Table 1. Cleavage, blastocyst rate, and MPF kinase activity in different sizes of prepubertal goat oocytes

scholarly journals Calophyllum brasilienseModulates the Immune Response and PromotesLeishmania amazonensisIntracellular Death

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
L. Domeneghetti ◽  
I. G. Demarchi ◽  
J. Z. Caitano ◽  
R. B. Pedroso ◽  
T. G. V. Silveira ◽  
...  
Keyword(s):  
Immune Response ◽  
Alternative Therapy ◽  
Folk Medicine ◽  
Rna Expression ◽  
Rt Pcr ◽  
Immunomodulatory Effects ◽  
Hydroalcoholic Extract ◽  
Calophyllum Brasiliense ◽  
Infection Index ◽  
Before And After

Calophyllum brasilienseis a plant from the Brazilian rain forests and has been used in folk medicine for the treatment of various diseases, including leishmaniasis. This infectious disease depends on theLeishmaniasp. and the host immune response.C. brasilienseantileishmanial activity is well known, but the effects on immune response remain to be investigated. This study showed the leishmanicidal and immunomodulatory effects of a 30 μg/mL of hydroalcoholic extract ofC. brasiliensein murine macrophages before and afterLeishmania(Leishmania)amazonensisinfection. The semiquantitative cytokine RNA expression was determined by RT-PCR and the anti-Leishmaniaactivity was measured by infection index (IF). Hydroalcoholic extract ofC. brasiliensereduced more than 95% of IF when used before and afterLeishmaniainfection, with 3 and 24 h of treatment (p<0.05).C. brasilienseinhibited or reduced significantly (p<0.05) the TNF-α, IL-1β, IL-18, and IL-10 mRNA expression. The antileishmanial and anti-inflammatory effects showed the potential ofC. brasilienseas an alternative therapy for leishmaniasis and it must be investigated.

RNA expression as a prognostic tool in idiopathic nephrotic syndrome

2007 ◽  
Vol 148 (23) ◽  
pp. 1067-1075
Author(s):  
Krisztina Fischer ◽  
Orsolya Galamb ◽  
Béla Molnár ◽  
Zsolt Tulassay ◽  
András Szabó
Keyword(s):  
Nephrotic Syndrome ◽  
Northern Blot ◽  
Idiopathic Nephrotic Syndrome ◽  
Minimal Change ◽  
Ribonuclease Protection Assay ◽  
Rna Expression ◽  
Rt Pcr ◽  
Protection Assay ◽  
Ribonuclease Protection

A gyermekkori nephrosis 90%-a idiopathiás nephrosis szindróma. Az idetartozó három kórkép, a minimal change betegség, a mesangialis proliferatio és a focalis sclerosis hasonló klinikai képpel jelentkező, eltérő prognózisú és terápiás válaszú betegség. Dolgozatunk célja az idiopathiás nephrosis szindrómába tartozó kórképek kialakulásával, progressziójával összefüggő genetikai ismeretek, génexpressziós változások áttekintése és funkcionális csoportosítása. A génexpressziós változások meghatározásának eszközeként, dolgozatunk röviden összefoglalja a northern blot, a ribonuclease protection assay, az in situ RNS-hibridizáció, a kvantitatív RT-PCR és a microarray módszerek lényegét. Az eddig elvégzett vizsgálatok a DNS-szintézis és repair gének, növekedési faktorok, extracelluláris mátrix, extracelluláris ligandreceptorok, extracelluláris jelátvitel zavarai mellett kiemelik a metabolikus és transzporter gének, illetve az immunszabályozó gének molekuláris eltéréseit, amelyek összefüggésben vannak az idiopathiás nephrosis szindróma eddig megismert molekuláris hátterével. A chiptechnológia fejlődésével és elterjedésével ezek a markerek és a hagyományos vizsgálati módszerek párhuzamos alkalmazása rutindiagnosztikai szempontból is fontossá válhat.

Changing trends in quality of media reporting of suicide in the community following a celebrity suicide in India

2021 ◽  
pp. 000486742110096
Author(s):  
Vikas Menon ◽  
Sujita Kumar Kar ◽  
Ramdas Ransing ◽  
Ginni Sharma ◽  
Jigyansa Ipsita Pattnaik ◽  
...  
Keyword(s):  
Online News ◽  
Significant Rise ◽  
Warning Signs ◽  
World Health ◽  
Media Reporting ◽  
Exact Test ◽  
Reporting Practices ◽  
Before And After ◽  
Celebrity Suicide

Objective: Little is known about changes in quality of media reporting of suicide in the community following a celebrity suicide. Our objective was to compare trends in quality of media reporting of suicide, before and after the suicide of an Indian entertainment celebrity, against the World Health Organization suicide reporting guidelines. Method: Online news portals of English and local language newspapers, as well as television channels, were searched to identify relevant suicide-related news articles. Comparison of reporting characteristics before and after the celebrity suicide was performed using chi-square test or Fisher’s exact test. Results: A total of 3867 eligible news reports were retrieved. There was a significant increase in harmful reporting characteristics, such as reporting the name, age and gender of the deceased ( p < 0.001 for all comparisons), mentioning the location ( p < 0.001) and reason for suicide ( p = 0.04) and including photos of the deceased ( p = 0.002) following the celebrity suicide. Helpful reporting practices were less affected; there was a significant rise in inclusion of expert opinion ( p = 0.04) and mention of suicide-related warning signs ( p = 0.02). Conclusion: Following a celebrity suicide, significant changes in the quality of media reporting of suicide were noted with an increase in several potentially harmful reporting characteristics.

scholarly journals Regulation of angiopoietin-like protein 4/fasting-induced adipose factor (Angptl4/FIAF) expression in mouse white adipose tissue and 3T3-L1 adipocytes

2008 ◽  
Vol 100 (1) ◽  
pp. 18-26 ◽  
Author(s):  
Sarah Dutton ◽  
Paul Trayhurn
Keyword(s):  
Skeletal Muscle ◽  
Cell Culture ◽  
Adipose Tissue ◽  
White Adipose Tissue ◽  
Culture Medium ◽  
Mrna Level ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Before And After ◽  
Stimulation Of

Angiopoietin-like protein 4 (Angptl4)/FIAF (fasting-induced adipose factor) was first identified as a target for PPAR and to be strongly induced in white adipose tissue (WAT) by fasting. Here we have examined the regulation of the expression and release of this adipokine in mouse WAT and in 3T3-L1 adipocytes. Angptl4/FIAF expression was measured by RT-PCR and real-time PCR; plasma Angptl4/FIAF and release of the protein in cell culture was determined by western blotting. The Angptl4/FIAF gene was expressed in each of the major WAT depots of mice, the mRNA level in WAT being similar to the liver and much higher (>50-fold) than skeletal muscle. Fasting mice (18 h) resulted in a substantial increase in Angptl4/FIAF mRNA in liver and muscle (9·5- and 21-fold, respectively); however, there was no effect of fasting on Angptl4/FIAF mRNA in WAT and the plasma level of Angptl4/FIAF was unchanged. The Angptl4/FIAF gene was expressed in 3T3-L1 adipocytes before and after differentiation, the level increasing post-differentiation; Angptl4/FIAF was released into the culture medium. Insulin, leptin, dexamethasone, noradrenaline, TNFα and several IL (IL-1β, IL-6, IL-10, IL-18) had little effect on Angptl4/FIAF mRNA levels in 3T3-L1 adipocytes. However, a major stimulation of Angptl4/FIAF expression was observed with rosiglitazone and the inflammatory prostaglandins PGD2 and PGJ2. Angptl4/FIAF does not act as an adipose tissue signal of nutritional status, but is markedly induced by fasting in liver and skeletal muscle.

Follicular progesterone concentrations and messenger RNA expression of MATER and OCT-4 in immature bovine oocytes as predictors of developmental competence

2015 ◽  
Vol 83 (7) ◽  
pp. 1179-1187 ◽  
Author(s):  
R. Urrego ◽  
E. Herrera-Puerta ◽  
N.A. Chavarria ◽  
O. Camargo ◽  
C. Wrenzycki ◽  
...  
Keyword(s):  
Messenger Rna ◽  
Developmental Competence ◽  
Rna Expression ◽  
Bovine Oocytes

scholarly journals Evidence-based practice of stereotactic radiosurgery: Outcomes from an educational course for neurosurgery and radiation oncology residents

2021 ◽  
Vol 12 ◽  
pp. 77
Author(s):  
Swathi Chidambaram ◽  
Sergio W. Guadix ◽  
John Kwon ◽  
Justin Tang ◽  
Amanda Rivera ◽  
...  
Keyword(s):  
Clinical Trials ◽  
Stereotactic Radiosurgery ◽  
Radiation Oncology ◽  
Evidence Based Practice ◽  
Evidence Base ◽  
Comfort Level ◽  
Evidence Based ◽  
Senior Faculty ◽  
Exact Test ◽  
Before And After

Background: As the field of brain and spine stereotactic radiosurgery (SRS) continues to grow, so will the need for a comprehensive evidence base. However, it is unclear to what degree trainees feel properly equipped to use SRS. We assess the perceptions and comfort level reported by neurosurgery and radiation oncology residents concerning the evidence-based practice of SRS. Methods: A continuing medical education (CME) course provided peer-reviewed updates regarding treatment with intracranial and spinal SRS. Presentations were given by neurosurgery and radiation oncology residents with mentorship by senior faculty. To gauge perceptions regarding SRS, attendees were surveyed. Responses before and after the course were analyzed using the Fisher’s exact test in R statistical software. Results: Participants reported the greatest knowledge improvements concerning data registries (P < 0.001) and clinical trials (P = 0.026). About 82% of all (n = 17) radiation oncology and neurosurgery residents either agreed or strongly agreed that a brain and spine SRS rotation would be beneficial in their training. However, only 47% agreed or strongly agreed that one was currently part of their training. In addition, knowledge gains in SRS indications (P = 0.084) and ability to seek collaboration with colleagues (P = 0.084) showed notable trends. Conclusion: There are clear knowledge gaps shared by potential future practitioners of SRS. Specifically, knowledge regarding SRS data registries, indications, and clinical trials offer potential areas for increased educational focus. Furthermore, the gap between enthusiasm for increased SRS training and the current availability of such training at medical institutions must be addressed.

scholarly journals Relationship Between Hand-washing Habit and Toilet Use with Diarrhea Incidence in Children Under Five Years

2017 ◽  
Vol 5 (1) ◽  
pp. 95
Author(s):  
Nikmatur Rohmah ◽  
Fariani Syahrul
Keyword(s):  
Public Health ◽  
Public Health Problem ◽  
Cross Sectional Study ◽  
Hand Washing ◽  
Chi Square ◽  
Cross Sectional ◽  
Exact Test ◽  
Children Under Five ◽  
Under Five ◽  
Before And After

Diarrhea is still a public health problem in the world and the mostly occur in children under five years. At the moment the number of diarrhea-caused mortality was 3.8 per 1000 per year by 3.2 episodes per year in children under five years. This research aims to analyze the association between hand-washing habits and toilet use with the incidence of diarrhea in children under five years. This is an observational analytic with cross sectional study. Population of the research was a children's mother took her children to the public health center of Sekardangan Sidoarjo Subdistrict. The number of samples taken as many as 58 mother of children under five years. Data collection was done with interviews to parents of children and the observations of the toilets. Technique of data analysis using chi-square test or Fisher's exact test. On the results of the research there is a significant association the habit of hand washing (p = 0.006) and toilet use(p = 0.014) with the incidenceof diarrhea in children under five years. Conclusions of the research is the habit of hand washing and toilet use had a significant association with the incidenceof diarrhea in children under five years. Advice that can be given to the mother of a children under five years is to wash the hands before and after defecating and before preparing food for children. Keywords: diarrhea, hand washing, toilet use, observational, children under five years

scholarly journals Rapid Point-Of-Care Serology and Clinical History Assessment Increase Protection Provided by RT-PCR Screening: A Pilot Study Involving Three Nursing Homes in Brescia, a Hotspot of Lombardy

2021 ◽  
Vol 9 ◽  
Author(s):  
Antonella Savio ◽  
Stefano Calza ◽  
Gianbattista Guerrini ◽  
Valentina Romano ◽  
Eleonora Marchina
Keyword(s):  
Nursing Homes ◽  
Close Contact ◽  
Fisher Exact Test ◽  
Rt Pcr ◽  
Positive Serology ◽  
Serological Screening ◽  
Pcr Screening ◽  
Exact Test ◽  
Viral Spread

Background: COVID-19 outbursts have been registered worldwide within care homes with asymptomatic transmission combined with shortage/inaccuracy of diagnostic tests undermining the efforts at containment of the disease. Nursing facilities in Lombardy (Italy) were left with no, or limited, access to testing for 8 weeks after the outbreak of COVID-19.Methods: This study includes 246 residents and 286 workers of three different nursing homes in Brescia-Lombardy. Clinical questionnaires and rapid serology tests were devised to integrate the data of the first available RT-PCR screening. Follow-up serology after 60-days was performed on 67 of 86 workers with positive serology or clinically suspicious.Findings: Thirty-seven residents and 18 workers had previous positive RT-PCR. Thorough screening disclosed two additional RT-PCR-positive workers. Serology screening revealed antibodies in 59 residents and 48 workers, including 32/37 residents and all workers previously positive at RT-PCR. Follow up serology disclosed antibodies in two additional workers with recent symptoms at the time of screening. The professionals in close contact with residents had more infections (47/226–20.79% vs. 1/60–1.66%; p = 0.00013 Fisher exact-test). A suspicious clinical score was present in 44/64 residents and in 41/50 workers who tested positive with either method with totally asymptomatic disease more frequent among residents 28.1 vs. 10.0% (p = 0.019 Fisher exact-test).Interpretation: Based on the available RT-PCR ± results at the time of symptoms/contacts, our integrated clinical and serological screening demonstrated sensitivity 89% and specificity 87%. This multimodal assessment proved extremely useful in understanding the viral spread in nursing homes, in defining its stage and in implementing protective measures. Rapid serology tests demonstrated efficient and particularly suited for older people less able to move/cooperate.

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